Adenosine Deaminase Activity Research Articles

Diagnostic values of adenosine deaminase cytokines in fluid contributed to the detection of extrapulmonary tuberculosis

Tuberculosis (TB) is a serious global public health problem. Detecting the changes of adenosine deaminase (ADA), interleukin (IL)-1β, IL-2, tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) released from macrophages and monocytes is greatly valuable in the diagnosis of TB. Hence, this study was designed to determine the optimal cut-off points of ADA, IL-1β, IL-2, TNF-α, and IFN-γ, allowing to differentiate pleural TB (PLTB), peritoneal TB (PTB), and meningeal TB (MTB). The study included 386 patients with pleural effusion, 47 patients with peritoneal effusion, and 134 patients with encephalitis and meningoencephalitis suspect of TB. ADA was determined by an enzyme kinetics method; cytokine concentration was measured by a test based on the competitive enzyme-linked immunosorbent assay principle. Sensitivity, specificity, and positive and negative predictive value were calculated and described. In the aspiration fluid, ADA activity was greatly different between PLTB, PTB, and MTB groups with values of 37.5 U/L, 30.5 U/L, and 8.1 U/L and sensitivity and specificity of 87.7% and 83.4%; 100% and 88.9%; 97.3% and 98.6%, respectively. Similar, IFN-γ concentrations were distinctively different between PLTB, PTB, and MTB patients (with values of 120 pg/mL, 200 pg/mL, and 30 pg/mL with sensitivity and specificity of 80.3% and 80.9%; 80.0% and 92.6%; 98.6% and 61.7%, respectively). The determination of ADA and IFN-γ levels in the aspiration fluid is potentially considered a highly sensitive and specific test for differentiating PLTB, PTB, and MTB.

The alleviating effects and mechanisms of Lactiplantibacillus plantarum MC14 on hyperuricemia in mice

Hyperuricemia (HUA) is a chronic disorder resulting from purine metabolic dysregulation. This research explored the alleviating effects and potential mechanisms of Lactiplantibacillus plantarum MC14 on HUA in a mouse model. Further, it investigated the impact of MC14 intervention on the intestinal flora and metabolites of HUA model mice. Results indicated that the activity of MC14 in alleviating HUA may be linked to its abilities to lower serum levels of uric acid, creatinine, and blood urea nitrogen, reduce inflammation, alleviate renal tissue damage, enhance uric acid excretion, inhibit the overactivation of the PI3K/AKT/mTOR signaling pathway, suppress hepatic adenosine deaminase and xanthine oxidase activities, competitively absorb nucleosides and nucleobases, and regulate the intestinal flora. Our research demonstrates that MC14 is a potential candidate for developing novel therapeutics for hyperuricemia.

Investigation of the Effect of Omega-3 Fatty Acids on Antioxidant System and Serum Aluminum, Zinc, and Iron Levels in Acute Aluminum Toxicity.

Aluminum (Al), one of the three most prevalent metals in the Earth's crust, adversely impacts all metabolic systems of living organisms due to its extensive utilization by humans. It is known that omega-3 fatty acids (ω-3FA) protect the organism against diseases and have positive effects on the immune system. The aim of the study was to investigate the effect of ω-3FA on 8-OH-2-deoxyguanosine (8-OHdG), glutathione (GSH) levels and adenosine deaminase (ADA), paraoxonase (PON), and catalase (CAT) activities in rats with acute aluminum toxicity. The study also aimed to investigate the antioxidant system, as well as Al, zinc (Zn), and iron (Fe) levels. Forty Sprague-Dawley rats (n = 40) were used in the study and the rats were divided into four equal groups (n = 10). In group I, 0.5mL of 0.9% saline solution (NaCI) was injected intraperitoneally. Group II was injected with 34mg/kg aluminum chloride (AlCI3) intraperitoneally. Group III received 400mg/kg ω-3FA for 7days and group IV received both AlCI3 and 400mg/kg ω-3FA for 7days. At the end of the study, blood samples were obtained by cardiac puncture. The findings showed that Al exposure increased serum 8-OHdG and total oxidant status (TOS) levels, as well as ADA activity, which are markers associated with oxidative damage. Conversely, PON and CAT activities, GSH, and total antioxidant status (TAS) levels decreased compared to the control group. Furthermore, Zn and Fe levels decreased as Al levels increased. In conclusion, Al has the capacity to induce oxidative damage and lipid peroxidation, while ω-3 fatty acids may mitigate this damage through a regulatory mechanism. Moreover, ω-3-FA could be used as a therapeutic agent that reduces Al toxicity.

State of Adenosine Deaminase in Patients with Dyslipidemia

ABSTRACT: Background: Dyslipidemia is becoming more common across all age groups mainly in young individuals because of imbalanced diets, low physical activity, and sedentary work culture. Several studies reported that elevated serum adenosine deaminase activity was associated with dyslipidemia, but the results were not consistent. Aim: It is aimed to correlate adenosine deaminase and lipid profile parameters in patients with dyslipidemia. Material and Methods: In this case-control study, a total of 60 subjects (30 diagnosed dyslipidemia patients and 30 age and gender-matched healthy individuals) were enrolled. Serum lipid profile parameters and adenosine deaminase levels were estimated in each subject. Results: The mean levels of lipid profiles, mainly triacylglycerol (TG), low-density lipoprotein-cholesterol (LDL-C), and adenosine deaminase, were found to be significantly high, while high-density lipoprotein-cholesterol (HDL-C) was found significantly low in cases than controls (p <0.001). adenosine deaminase has not shown any significant correlation with lipid profile parameters in patients with dyslipidemia and controls. Conclusion: The result showed that the serum adenosine deaminase and serum lipid profile levels were altered in patients with dyslipidemia.

Adenosine deaminase is a risk factor for mortality after discharge in patients with acute myocardial infarction: Long-term clinical follow-up

BackgroundVariations in adenosine deaminase (ADA) activity have been detected in numerous cardiovascular diseases (CVDs), but there is limited research on its role in the prognosis of CVDs. In this study, we explored the role of ADA in the prognosis of patients with acute myocardial infarction (AMI). MethodIn this study, a total of 1,574 patients with a first diagnosis of acute myocardial infarction (AMI) were followed up for a median (interquartile range [IQR]) of 77.0 (50.0, 95.0) months after discharge. Cox proportional hazards regression models were used to identify factors that are substantially valuable for patient prognosis. ResultsDuring the follow-up period, the mortality rate of AMI was 12.5 %. The 3-year and 5-year overall survival (OS) rates of AMI patients were 93.8 % and 91.0 %, respectively. Multivariate Cox regression analysis revealed that serum ADA (hazard ratio [HR] = 1.166, 95 % confidence interval [CI]: 1.006–1.352) was an independent risk factor for 5-year OS after discharge in AMI patients. When serum ADA was assessed in quartiles, compared with the reference group (Quartile 1), the adjusted HR for death was 2.498 (95 % CI: 1.344–4.642) in Quartile 4 for 5-year OS and 2.508 (95 % CI: 1.145–5.496) in Quartile 4 for 3-year OS. ConclusionsSerum ADA levels at admission are a risk factor that affects the long-term prognosis of AMI patients after hospital discharge.

Anti-inflammatory potential of polyphenols: Combining in silico prediction and in vivo data

Introduction: Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by joint inflammation and destruction, leading to significant pain and disability. Adenosine deaminase (ADA) is identified as a biomarker for RA’s inflammatory process. This study aims to investigate the potential of flavonoids and phenolic acids to inhibit ADA activity (in silico) and evaluate their anti-inflammatory effects in a RA model (in vivo). Methods: The molecular docking study was conducted using YASARA Structure 19.12.14. software following the Auto Dock 4.2 protocol. A rat model with pristane-induced arthritis was used to test the anti-inflammatory effect of selected polyphenols. The consistency of the development of the rat model was evaluated through the following indicators artistic score, paw volume, and body weight. Quercetin was administered intragastrically at doses of 150 and 400 mg/kg over 15 days. The C-reactive protein (CRP) level in serum was measured with an automatic biochemical analyzer. Statistical analyses were performed using SPSS 29.0.2.0. Results: Molecular docking simulations showed flavonoids inhibited ADA activity with inhibition constants ranging from 0.012 mM to 0.190 mM. In the in vivo RA model, quercetin significantly reduced joint inflammation and serum CRP levels at a higher dose of 400 mg/kg. Conclusion: Quercetin shows promise as an anti-inflammatory agent for RA by targeting ADA, suggesting that flavonoid-rich plant extracts could enhance RA treatment.

Enzyme-responsive aptasensing based on the ATP aptamer and benzothiazole-coumarin G-quadruplex fluorescent probe for label-free detection of adenosine deaminase activity and its inhibitor

Adenosine deaminase (ADA) is a vital enzyme for regulating the biological process in humans and its abnormal expression level is related to various severe diseases. Nevertheless, conventional approaches for detecting ADA activity are hindered by the drawbacks of complexity and limited detection performances. To address these challenges, we here designed an ADA-responsive aptasensing strategy based on a synthesized benzothiazole-coumarin G-quadruplex (G4) fluorescent probe, which can bind to the G-rich ATP aptamer (G-APA) and form the high-contrast fluorescent complex (S1/G-APA). While the fluorescence of S1/G-APA could be quenched by ATP competition due to the allosteric process, the deamination in ATP catalyzed by ADA would recover the fluorescence of S1/G-APA and allow a facile, turn-on, label-free, and selective detection of ADA activity. This proposed method exhibits favorable detection performances for ADA activity analysis with a wide detection linear range varied from 1 to 100 U/L and a detection limit of 0.82 U/L. Meanwhile, it can be applied for ADA inhibitor (DCF) evaluation and provide a turn-off and sensitive detection performances. Inspired by these detection efficacies, the serum samples spiked with ADA and DCF have been investigated to present the satisfied recovery rates and detection reliability. Collectively, this method will provide a promising way for portable and efficient ADA activity quantification and evaluating the inhibition efficiencies of ADA inhibitors, which hold the potential in promoting ADA-related sensing and pharmaceutical research.

Engineering TadA ortholog-derived cytosine base editor without motif preference and adenosine activity limitation

The engineered TadA variants used in cytosine base editors (CBEs) present distinctive advantages, including a smaller size and fewer off-target effects compared to cytosine base editors that rely on natural deaminases. However, the current TadA variants demonstrate a preference for base editing in DNA with specific motif sequences and possess dual deaminase activity, acting on both cytosine and adenosine in adjacent positions, limiting their application scope. To address these issues, we employ TadA orthologs screening and multi sequence alignment (MSA)-guided protein engineering techniques to create a highly effective cytosine base editor (aTdCBE) without motif and adenosine deaminase activity limitations. Notably, the delivery of aTdCBE to a humanized mouse model of Duchenne muscular dystrophy (DMD) mice achieves robust exon 55 skipping and restoration of dystrophin expression. Our advancement in engineering TadA ortholog for cytosine editing enriches the base editing toolkits for gene-editing therapy and other potential applications.

Effects and mechanisms of Polygonati Rhizoma polysaccharide on potassium oxonate and hypoxanthine-induced hyperuricemia in mice

Hyperuricemia, a prevalent metabolic disturbance intricately linked to gout and chronic kidney disease (CKD), may be relieved by traditional Chinese medicine Polygonati Rhizoma. It is derived from the rhizomes of Polygonatum sibiricum, Polygonatum kingianum, and Polygonatum cyrtonema, which are rich in polysaccharides and are effective hyperuricemia alleviators. This study investigated the potential of Polygonatum sibiricum polysaccharide (PSP) in managing hyperuricemia. PSP (125, 250, and 500 mg/kg, i.g.) or allopurinol was administered to hyperuricemia mice treated with potassium oxonate and hypoxanthine for two weeks. PSP effectively decreased serum uric acid levels by inhibiting xanthine oxidase and adenosine deaminase activity and expression in the liver and modulating uric acid-related transporters (URAT1, OAT1, and OAT3) in the kidney. PSP lowered serum creatinine and blood urea nitrogen levels, alleviating hyperuricemia-induced renal tubular epithelial-mesenchymal fibrosis. In vitro, PSP promoted mitochondrial biogenesis via the PGC-1α/NRF1/TFAM pathway, suppressed reactive oxygen species production, and prevented cytochrome C and dynamin-related protein 1 dysregulation in HK-2 cells. Furthermore, PSPA (Mw 4.0 kDa) and PSPB (Mw 112.2 kDa) isolated from PSP exhibit different uric acid-lowering mechanisms. In conclusion, our findings highlight the therapeutic potential of PSP and its nephroprotective effects in hyperuricemia, thereby supporting its development as a therapeutic agent for hyperuricemia.

Kinetic Determination of Cerebrospinal Fluid Adenosine Deaminase Activity for the Diagnosis of Tuberculous Meningitis.

Tuberculous meningitis (TBM) is a common infectious disease of the central nervous system. Detection of cerebrospinal fluid (CSF) adenosine deaminase (ADA) activity for the diagnosis of TBM has a relatively high accuracy. Most previous reports determined ADA levels by following the colorimetric method of Giusti (endpoint assay). Recently, a Diazyme ADA assay kit was developed to determine CSF ADA levels. This study aimed to define the cutoff value of CSF ADA that is diagnostic for TBM with the Diazyme ADA assay kit. Adults with meningitis were included in the study. The diagnostic properties of CSF ADA for TBM as determined by the Diazyme ADA assay kit were assessed by the receiver operating characteristic curve, area under the curve, sensitivity, specificity, and likelihood ratios. There were 97 patients enrolled in the study, comprising 15 cases of TBM and 82 cases of non-TBM meningitis. The median CSF ADA activity level in the TBM group was significantly higher than in the non-TBM group (P = 0.002), with a mean difference of 14.5 U/L (95% CI: 5.3-23.8 U/L). A CSF ADA level of 6.1 U/L was the best cutoff value to differentiate between the TBM and non-TBM groups, with a sensitivity of 53.33%, a specificity of 89.02%, a positive likelihood ratio of 4.86, a negative likelihood ratio of 0.52, and an area under the curve of 0.75. A CSF ADA level of 6.1 U/L determined by the Diazyme ADA assay kit could be used as a diagnostic tool in the early diagnosis of TBM.

Mechanisms of levan in ameliorating hyperuricemia: Insight into levan on serum metabolites, gut microbiota, and function in hyperuricemia rats

This study aims to investigate the effects of levan on the progression of hyperuricemia (HUA) rats and elucidate its underlying mechanisms. After levan intervention, both low and high-dose groups exhibited a significant decrease in serum uric acid (UA) levels, reaching 71.0 % and 77.5 %, respectively, compared to the model group. Furthermore, levan could alleviate renal pathological damage caused by glomerular cell vacuolation, inflammatory infiltration and collagen deposition. The results of enzyme activity assay and real-time fluorescence quantitative PCR showed that levan decreased UA production by inhibiting adenosine deaminase (ADA) activity and gene expression in liver; it upregulated ATP-binding cassette subfamily G member 2 protein (ABCG2) and organic anion transporter 1 (OAT1) transporter gene expression in the kidney, promoting UA excretion. Gut microbiome analysis indicated that levan regulated gut flora dysbiosis induced by HUA, resulting in up-regulated the abundance of beneficial bacteria (Muribaculaceae, Faecalibaculum, Bifidobacterium, and Lactobacillus) and decreased conditioned pathogenic bacteria (Escherichia_Shigella and Proteus). Non-targeted metabolomics showed changes in various serum metabolites associated with glycerophospholipid metabolism, lipid metabolism, and inflammation following oral administration of levan. Therefore, levan may be a promising functional dietary supplement for regulating the gut flora and remodeling of metabolic disorders in individuals with HUA.

Evolving spectrum of adenosine deaminase (ADA) deficiency: Assessing genotype pathogenicity according to expressed ADA activity of 46 variants

Deficiency of adenosine deaminase (ADA or ADA1) has broad clinical and genetic heterogeneity. Screening techniques can identify asymptomatic infants whose phenotype and prognosis are indeterminate, and who may carry ADA variants of unknown significance. We systematically assessed the pathogenic potential of rare ADA missense variants to better define the relationship of genotype to red blood cell (RBC) total deoxyadenosine nucleotide (dAXP) content and to phenotype. We expressed 46 ADA missense variants in the ADA-deficient SØ3834 strain of Escherichia coli and defined genotype categories (GCs) ranked I to IV by increasing expressed ADA activity. We assessed relationships among GC rank, RBC dAXP, and phenotype in 58 reference patients with 50 different genotypes. We used our GC ranking system to benchmark AlphaMissense for predicting variant pathogenicity, and we used a minigene assay to identify exonic splicing variants in ADA exon 9. The 46 missense variants expressed ∼0.001% to ∼70% of wild-type ADA activity (40% had <0.05% of wild-type ADA activity and 50% expressed >1%). RBC dAXP ranged from undetectable to >75% of total adenine nucleotides and correlated well with phenotype. Both RBC dAXP and clinical severity were inversely related to total ADA activity expressed by both inherited variants. Our GC scoring system performed better than AlphaMissense in assessing variant pathogenicity, particularly for less deleterious variants. For ADA deficiency, pathogenicity is a continuum and conditional, depending on the total ADA activity contributed by both inherited variants as indicated by GC rank. However, in patients with indeterminate phenotype identified by screening, RBC dAXP measured at diagnosis may have greater prognostic value than GC rank.

Effect of curcumin-donepezil combination on spatial memory, astrocyte activation, and cholinesterase expressions in brain of scopolamine-treated rats.

The study investigated the effect of co-administration of curcumin and donepezil on several markers of cognitive function (such as spatial memory, astrocyte activation, cholinesterase expressions) in the brain cortex and hippocampus of scopolamine-treated rats. For seven consecutive days, a pre-treatment of curcumin (50mg/kg) and/or donepezil (2.5mg/kg) was administered. On the seventh day, scopolamine (1mg/kg) was administered to elicit cognitive impairment, 30min before memory test was conducted. This was followed by evaluating changes in spatial memory, cholinesterase, and adenosine deaminase (ADA) activities, as well as nitric oxide (NO) level were determined. Additionally, RT-qPCR for glial fibrillary acidic protein (GFAP) and cholinesterase gene expressions was performed in the brain cortex and hippocampus. Also, GFAP immunohistochemistry of the brain tissues for neuronal injury were performed in the brain cortex and hippocampus. In comparison to the control group, rats given scopolamine had impaired memory, higher levels of acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and ADA activities, as well as elevated markers of oxidative stress. In addition to enhanced GFAP immunoreactivity, there was also overexpression of the GFAP and BChE genes in the brain tissues. The combination of curcumin and donepezil was, however, observed tobetter ameliorate these impairments in comparison to thedonepezil-administered rat group. Hence, this evidence provides more mechanisms to support the hypothesis that the concurrent administration of curcumin and donepezil mitigates markers of cognitive dysfunction in scopolamine-treated rat model.

9-Hydroxy-8-oxypalmatine, a novel liver-mediated oxymetabolite of palmatine, alleviates hyperuricemia and kidney inflammation in hyperuricemic mice

Ethnopharmacological relevancePalmatine is a main bioactive alkaloid of Cortex Phellodendri, which has been commonly prescribed for the treatment of hyperuricemia (HUA) in China. The metabolites of palmatine were crucial to its prominent biological activity. 9-Hydroxy-8-oxypalmatine (9-OPAL) is a novel liver-mediated secondary oxymetabolite of palmatine. Aim of the studyThe current study was to assess the efficacy of 9-OPAL, a novel liver-mediated secondary oxymetabolite of palmatine derived from Cortex Phellodendri, in experimental HUA mouse model and further explore its underlying mechanism. Materials and methodsAn in vitro metabolic experiment with oxypalmatine was carried out using liver samples. We separated and identified a novel liver metabolite, and investigated its anti-HUA effect in mice. HUA mice were induced by potassium oxonate and hypoxanthine daily for one week. After 1 h of modeling, mice were orally administered with different doses of 9-OPAL (5, 10 and 20 mg/kg). The pathological changes of the kidneys were evaluated using hematoxylin-eosin staining (H&E). The acute toxicity of 9-OPAL was assessed. The effects of 9-OPAL on serum levels of uric acid (UA), adenosine deaminase (ADA), xanthine oxidase (XOD), creatinine (CRE), blood urea nitrogen (BUN) and inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA) or biochemical method. Furthermore, Western blot, quantitative real-time PCR (qRT-PCR) and molecular docking were used to investigate the effect of 9-OPAL on the expression of renal urate transporters and NLRP3 signaling pathway in HUA mice. Results9-OPAL had been discovered to be a novel liver-mediated oxymetabolite of palmatine for the first time. Treatment with 9-OPAL significantly reduced the UA, CRE as well as BUN levels, and also effectively attenuated abnormal renal histopathological deterioration with favorable safety profile. Besides, 9-OPAL significantly decreased the serum and hepatic activities of XOD and ADA, dramatically inhibited the up-regulation of UA transporter protein 1 (URAT1) and glucose transporter protein 9 (GLUT9), and reversed the down-regulation of organic anion transporter protein 1 (OAT1). Additionally, 9-OPAL effectively mitigated the renal inflammatory markers (TNF-α, IL-1β, IL-6 and IL-18), and downregulated the transcriptional and translational expressions of renal Nod-like receptor family pyrin domain containing 3 (NLRP3), caspase-1, apoptosis-associated speck-like (ASC) and IL-1β in HUA mice. Molecular docking results revealed 9-OPAL bound firmly with XOD, OAT1, GLUT9, URAT1, NLRP3, caspase-1, ASC and IL-1β. Conclusions9-OPAL was found to be a novel liver-mediated secondary metabolite of palmatine with favorable safety profile. 9-OPAL had eminent anti-hyperuricemic and renal-protective effects, and the mechanisms might be intimately associated with repressing XOD activities, modulating renal urate transporter expression and suppressing the NLRP3 inflammasome activation. Our investigation might also provide further experimental evidence for the traditional application of Cortex Phellodendri in the treatment of HUA.

To Study the Combined Use of Pleural Fluid Lymphocyte / Neutrophil Ratio and ADA for the Diagnosis of Tuberculous Pleural Effusion

INTRODUCTION - Pleural effusion is the abnormal collection of fluid in the pleural space. It is classified into exudates and transudates based on Light's criteria. Among the extra pulmonary presentations after tuberculous lymphadenitis, pleural TB is the second frequent. Although many biochemical parameters, such as lactate dehydrogenase (LDH), C-reactive protein (CRP), adenosine deaminase (ADA), interferon-gamma, and procalcitonin levels have been studied in the context of the diagnosis of exudative pleural effusion, its diagnosis is still challenging. AIMS &amp; OBJECTIVES- To determine whether the combined use of ADA activity and lymphocyte/neutrophil ratio would provide a more efficient means for diagnosing tuberculous pleural effusion than the use of ADA alone. MATERIAL &amp; METHODS- Hospital based Observational study was conducted over 8 months with total number of 100 Patients diagnosed as having pleural effusion based on clinical features and chest radiography in GEMS MEDICAL COLLEGE AND HOSPITAL. Biochemical, cytological and microbiology studies were done by obtaining pleural fluid by thoracocentesis. RESULTS - out of 100 cases, 84 cases were tubercular and had high level of ADA in comparison to rest of 16 non tubercular cases. At level of 50 IU/L of ADA activity test had sensitivity of 97.6%, specificity 87.5% which was increased to 100% and 92.8% in combination with test of Lymphocytic/Neutrophilic ratio - 0.75 CONCLUSION- The combined use of ADA along with the L/N ratio would provide a more efficient means for diagnosing tuberculous pleural effusion than the use of ADA alone.

Biochemical risk factors associated with refractory epilepsy: alpha synuclein and adenosine deaminase

Abstract Background Epilepsy is a common chronic neurological disorder affecting all age groups. A significant portion of children with epilepsy develop drug-resistant seizures. These children are at risk of cognitive and behavioral comorbidities and death. Some clinical features provide important information about the prognosis of epilepsy. However, currently, there is no objective biochemical indicator associated with refractory epilepsy. This study aimed to determine whether serum alpha-synuclein (αS), pglycoprotein (P-gp), and adenosine deaminase activity (ADA) were biochemical risk factors for refractory epilepsy. Methods The cross-sectional study included patients diagnosed with refractory epilepsy (n=32), non-refractory epilepsy (n=35) and 20 healthy children under the age of 18 who applied to the Pediatric Neurology outpatient clinic. In the serum samples, αS and P-gp were analyzed by ELISA and ADA activity was analyzed by spectrophotometric method. ROC analysis was applied and the area under the curve (AUC) was calculated to define cut-off values in distinguishing refractory epilepsy patients from non-refractory epilepsy patients. Binary logistic regression analysis was performed to define risk factors associated with resistance in patients with epilepsy. Results Children with epilepsy whose ADA activity and αS values were higher than the determined cut-off values had 10-fold and 5.3-fold increased risk of refractory epilepsy, respectively. Conclusions αS and ADA activity can be used as biochemical risk factors for refractory epilepsy. However, these results need to be confirmed by prospective studies with many patients.

Intermittent high altitude hypoxia induced liver and kidney injury leading to hyperuricemia

About 140 million people worldwide live at an altitude above 2500 m. Studies have showed an increase of the incidence of hyperuricemia among plateau populations, but little is known about the possible mechanisms. This study aims to assess the effects of high altitude on hyperuricemia and explore the corresponding mechanisms at the histological, inflammatory and molecular levels. This study finds that intermittent hypobaric hypoxia (IHH) exposure results in an increase of serum uric acid level and a decrease of uric acid clearance rate. Compared with the control group, the IHH group shows significant increases in hemoglobin concentration (HGB) and red blood cell counts (RBC), indicating that high altitude hyperuricemia is associated with polycythemia. This study also shows that IHH exposure induces oxidative stress, which causes the injury of liver and renal structures and functions. Additionally, altered expressions of organic anion transporter 1 (OAT1) and organic cation transporter 1 (OCT1) of kidney have been detected in the IHH exposed rats. The adenosine deaminase (ADA) expression levels and the xanthione oxidase (XOD) and ADA activity of liver of the IHH exposure group have significantly increased compared with those of the control group. Furthermore, the spleen coefficients, IL-2, IL-1β and IL-8, have seen significant increases among the IHH exposure group. TLR/MyD88/NF-κB pathway is activated in the process of IHH induced inflammatory response in joints. Importantly, these results jointly show that IHH exposure causes hyperuricemia. IHH induced oxidative stress along with liver and kidney injury, unusual expression of the uric acid synthesis/excretion regulator and inflammatory response, thus suggesting a potential mechanism underlying IHH-induced hyperuricemia.

Effect of Specific Spoilage Organisms on the Degradation of ATP-Related Compounds in Vacuum-Packed Refrigerated Large Yellow Croaker (Larimichthys crocea).

This study examined the spoilage potential of specific spoilage organisms on the degradation of adenosine triphosphate (ATP)-related compounds in vacuum-packed refrigerated large yellow croaker. The total viable count (TVC), ATP-related compounds and related enzymes of vacuum-packed refrigerated large yellow croaker inoculated with different bacteria (Pseudomonas fluorescens and Shewanella putrefaciens) were characterized using the spread plate method, high-performance liquid chromatography and assay kits, respectively. Results indicated that the TVC for both control and Shewanella putrefaciens groups reached spoilage levels at days 9 and 15, respectively. The changes of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP) and adenosine deaminase activity across all groups showed no significant difference attributable to microbial growth. The results suggested that ATP to inosine monophosphate (IMP) degradation primarily occurs via fish's endogenous enzymes, with minimal microbial involvement. On day 12, the IMP content in fillets inoculated with Pseudomonas fluorescens (0.93 μmol/g) was half higher than in those inoculated with Shewanella putrefaciens (0.57 μmol/g). Both spoilage organisms facilitated IMP degradation, with Shewanella putrefaciens making a more substantial contribution. Analysis of K values and correlation coefficients revealed that Shewanella putrefaciens was the primary factor in the freshness loss of refrigerated vacuum-packed large yellow croaker. These findings offer a reference for understanding quality changes in refrigerated large yellow croaker, especially regarding umami degradation at the microbial level.

NCas9 Engineering for Improved Target Interaction Presents an Effective Strategy to Enhance Base Editing.

Base editors (BEs) are a recent generation of genome editing tools that couple a cytidine or adenosine deaminase activity to a catalytically impaired Cas9 moiety (nCas9) to enable specific base conversions at the targeted genomic loci. Given their strong application potential, BEs are under active developments toward greater levels of efficiency and safety. Here, a previously overlooked nCas9-centric strategy is explored for enhancement of BE. Based on a cytosine BE (CBE), 20 point mutations associated with nCas9-target interaction are tested. Subsequently, from the initial positive X-to-arginine hits, combinatorial modifications are applied to establish further enhanced CBE variants (1.1-1.3). Parallel nCas9 modifications in other versions of CBEs including A3A-Y130F-BE4max, YEE-BE4max, CGBE, and split-AncBE4max, as well as in the context of two adenine BEs (ABE), likewise enhance their respective activities. The same strategy also substantially improves the efficiencies of high-fidelity nCas9/BEs. Further evidence confirms that the stabilization of nCas9-substrate interactions underlies the enhanced BE activities. In support of their translational potential, the engineered CBE and ABE variants respectively enable 82% and 25% higher rates of editing than the controls in primary human T-cells. This study thus demonstrates a highly adaptable strategy for enhancing BE, and for optimizing other forms of Cas9-derived tools.

Clinical Significance of Serum ADA Combined with GLB, CREA, β2-MG and HGB in Newly Diagnosed Multiple Myeloma

To investigate the role of serum adenosine deaminase (ADA) combined with globulin (GLB), creatinine (CREA), β2-microglobulin (β2-MG) and hemoglobin (HGB) in the initial screening of multiple myeloma (MM), in order to reduce missed diagnosis and misdiagnosis of MM. A retrospective analysis was performed on 62 newly diagnosed multiple myeloma (NDMM) patients who were admitted to the Department of Hematology of the First Affiliated Hospital of Chengdu Medical College from April 2018 to December 2021, and 33 patients with benign hematologic diseases and 30 healthy subjects were selected as the control group. The expression of ADA in pan-cancer was analyzed using TCGA and GTEx databases. The general data and laboratory indicators of the subjects were collected, and the differences of ADA activity and other laboratory indicators in each group were compared. The relationship between serum ADA activity and clinical data of NDMM patients was analyzed. The changes of ADA activity before and after chemotherapy in NDMM patients and the differences of ADA activity in NDMM patients with different DS and ISS stages were compared. Multivariate logistic regression was used to analyze the risk factors of NDMM. The receiver operating characteristic(ROC) curve was used to evaluate the diagnostic efficacy of ADA and other laboratory indicators in MM. Bioinformatics method was used to analyze the co-expression networks and enrichment pathways of ADA. ADA level was significantly upregulated in tissues of 14 types of cancer in TCGA database, and ADA was highly expressed in 11 types of cancer in TCGA combined with GTEx databases. The serum levels of ADA, GLB, uric acid (UA), cystatin C (CysC) and β2-MG in the NDMM group were significantly higher than those in benign hematologic disease group and healthy control group ( P < 0.05), while the levels of ALB and the value of albumin to globulin ratio (A∶G) in the NDMM group were significantly lower than those in the other two groups ( P < 0.001). There were significant differences in DS stage (P =0.036), ISS stage (P =0.019) and the levels of CREA (P =0.036), UA (P =0.034), β2-MG (P =0.019) in NDMM patients with different ADA activity levels. After primary chemotherapy, ADA activity and β2-MG concentration were decreased in NDMM patients ( P < 0.01). The comparison results of patients in different stages showed that ADA activity of patients in DS stage I+II was significantly lower than that of patients in DS stage III (P <0.05), and ADA activity of patiens in ISS stage I+II was significantly lower than that of patients in ISS stage III ( P < 0.01). Multivariate logistic regression analysis showed that increased GLB, increased ADA activity, increased CREA, increased β2-MG and decreased HGB were independent risk factors for NDMM. The area under the curve (AUC) of ADA in the diagnosis of MM was 0.847, and the AUC of ADA combined with GLB, CREA, β2-MG and HGB in the diagnosis of MM was 0.940. The results of co-expression network and enrichment pathway analysis showed that ADA bounded to 20 proteins and it was significantly associated with the metabolic pathways of purine, pyrimidine, nicotinate and nicotinamide. The detection of ADA activity in serum is of positive significance for the auxiliary diagnosis, therapeutic evaluation and monitoring the progress of NDMM patients. ADA combined with GLB, CREA, β2-MG and HGB can improve the detection rate of MM, and reduce missed diagnosis and misdiagnosis to a certain extent.